Despite the advances in medical research, diagnostic testing strategies in current use often are not cost effective, are time-intensive, are limited in the scope of antigen tested for, and/or are labor-intensive. There is a great need for additional diagnostic methods and devices capable of detecting or assessing the health or identifying or indicating the presence of disease or disease-causing factors in humans and animals.
Diagnostic testing methods are needed for identifying or indicating the presence of various diseases, infections, and conditions.
The mammalian immune system reacts to foreign agents such as organisms and allergens through numerous recognition pathways involving the innate and adaptive immune systems. Normally, the immune system is capable of limiting or eliminating infection by a foreign organism, however, some organisms have mechanisms to evade the human immune system and can cause disease, morbidity and death. Similarly, the immune system typically generates an immune response against allografts or xenografts in organ and tissue transplant recipients causing rejection of the transplant. Further, in an increasing number of cases, the immune system is hyperactive, and reacts to self or environmental agents that it would normally be tolerant to, causing autoimmune, asthmatic or allergic responses. The aforementioned acute or chronic inflammatory states caused by infection, autoimmune and allergic diseases are significant causes of morbidity and mortality worldwide. Infectious diseases (ID) are clinically and sub-clinically manifested diseases caused by the presence of microbial organisms including pathogenic bacteria, fungi, protozoa, parasites and agents such as viruses and prions. ID are communicable diseases spread through sexual or casual contact, food, water and air. Further, infectious diseases cause significant loss to the global economy, but more importantly, they account for 16% of all deaths annually, 55% of deaths in children under the age of five years, and 26% of deaths in neonates. Many of the common IDs are largely preventable and are easy to treat once identified.
For example, rapid identification of methicillin-resistant S. aureus (MRSA) is highly desirable, as it is the leading cause of hospital-acquired infections (HAI). There is a need for more timely results on MRSA patient screening in clinical settings. Current technologies that exist for MRSA detection are PCR, FISH, lateral flow immunoassay, phage infection, latex agglutination or plating on selective media for MRSA sample screening; each requiring hours of incubation and sample preparation. Limitations of exclusively nucleic acid based approaches (PCR and FISH) are that multiple mutations in the target region, SCCmec, have been determined and decreased test sensitivity can result due to low probe hybridization efficiency. Limitations of traditional plating is low sensitivity and long time-to-answer. Limitations of the latex agglutination assay is the ambiguity of a positive test result, as the test must be read within a small timeframe and is not practical for many settings. Limitations of the phage-based tests require metabolically active intact cells. Limitations of current lateral flow immunoassays are the requirements to culture the organism and to prepare a sample and the high limits of detection (due to the binding of a single epitope on an analyte). Net-based assays for the detection of MRSA analytes enable multiple signatures (nucleic acid and protein) to be detected simultaneously and in a highly specific manner (refer to data on the lack of non-specific binding). The molecular net does not require sample preparation and is constructed to recognize multiple epitopes on a single analyte.
In addition, infections acquired in the community results in over 12 million ambulatory care visits/year, and are the main cause of skin and soft tissue infections in emergency departments. Currently, MRSA strain testing takes hours or days, which is too late to guide critical decisions about admission, isolation, and treatment. Additionally, acute and prolonged infections can result in an overly robust immune response leading to sepsis (septicemia), which is a serious medical condition and is the tenth leading cause of death in the US (CDC, 2003). Populations at risk of septicemia are the elderly, immunocompromised and critically ill patients. Sepsis is usually treated with intravenous fluids and antibiotics, however insufficient or delayed treatment can lead to severe sepsis characterized by organ dysfunction, hypotension, multi-organ failure and shock. Mortality rates for severe sepsis are over 60% in the United States each year. Allergies and food intolerances arise from hypersensitivity reactions of the immune system to one or more allergens. Allergic diseases affect as many as 50 to 60 million Americans and its prevalence is increasing. Food allergies account for 35-50% of all cases of anaphylaxis. More than 54% of all United States citizens are estimated to have one or more allergy (CDC). The prevalence of allergic rhinitis is approximately 35% in Europe and Australia (ECRHS). The number of ambulatory care visits in the United States with the primary diagnosis of allergic rhinitis is over 13 million in 2008. From 2000-2005, the cost of treating allergic rhinitis is increasing from $6.1 billion to $11.2 billion with more than half of the money spent on prescription medications. An allergen must be exposed to an organism to elicit a response, usually upon inhalation or ingestion. An allergen is a molecule, which causes an abnormal immunological response. Allergens tend to be protein but, can also be smaller and are called haptens.
Allergens can be bound by IgE immunoglobulins, B lymphocytes, T lymphocytes and Mast cells in the human. IgE-allergen immune complexes cause de-granulation of Mast cells and a release of histamines, locally and systemically. Histamine release can cause muscle cramps, muscle contraction, inflammation, erythema and edema of mucous membranes. Severe reactions can cause anaphylaxis and death.
Metabolic diseases (MD) comprise a large class of diseases involving disorders in metabolism. Many MD have a genetic component in addition to environmental components. Many of the genetically induced MD are due to genetic defects in enzyme function. Most MD arise due to the accumulation of enzyme substrates or enzyme products, the buildup of which is cytotoxic.
Lifestyle diseases (LD), sometimes called diseases of longevity, are diseases that appear in industrialized populations and populations with increased lifespan. Some common LD are Alzheimer's disease, atherosclerosis, hypertension, asthma, cancer, chronic liver disease, chronic obstructive pulmonary disease, Type 2 diabetes, heart disease, nephritis, osteoporosis, acne, stroke, depression, attention deficit and hyperactivity disorder, and obesity. Since the late 1990's, LD have accounted for over 60 percent of all mortality (National Center for Health Statistics, National Office of Vital Statistics).
Metabolic diseases, chronic diseases, cancers, autoimmune diseases, neuronal disorders and diseases, age-related diseases and infectious diseases are usually assessed by iterative testing strategies involving traditional physiologic measurements in the clinic, and more recently molecular assessments and measurements of molecules that are indicative of specific diseases in clinical labs. Despite the advances in medical research, there exists great need for additional diagnostics capable of detecting or assessing the health or identifying or indicating the presence of disease or disease-causing factors or disease-modifying factors in mammals. Accordingly, it is an objective of the present invention to utilize a device composing a molecular net on a polymeric platform or a kit of devices to be used in the diagnosis, detection or indication of specific aspects of health, disease, and disease-causing agents in mammals. Current diagnostic testing strategies are not cost effective, are time-intensive, are limited in the scope of antigen tested for, and/or are labor-intensive. Fast knowledge of individual health status allows for (i) appropriate medical treatment, (ii) the modification of behavior to decrease the likelihood disease incidence, morbidity, infection propagation and, (iii) the ability to save lives.
The mammalian immune system reacts to foreign agents such as organisms and allergens through numerous recognition pathways involving the innate and adaptive immune systems. Normally, the immune system is capable of limiting or eliminating infection by a foreign organism, however, some organisms have mechanisms to evade the human immune system and can cause disease, morbidity and death. Similarly, the immune system typically generates an immune response against allografts or xenografts in organ and tissue transplant recipients causing rejection of the transplant. Further, in an increasing number of cases, the immune system is hyperactive, and reacts to self or environmental agents that it would normally be tolerant to, causing autoimmune, asthmatic or allergic responses. The aforementioned acute or chronic inflammatory states caused by infection, autoimmune and allergic diseases are significant causes of morbidity and mortality worldwide.
Allergies and food intolerances arise from hypersensitivity reactions of the immune system to one or more allergens. Allergic diseases affect as many as 50 to 60 million Americans and its prevalence is increasing. Food allergies account for 35-50% of all cases of anaphylaxis. More than 54% of all United States citizens are estimated to have one or more allergy (CDC). The prevalence of allergic rhinitis is approximately 35% in Europe and Australia (ECRHS). The number of ambulatory care visits in the United States with the primary diagnosis of allergic rhinitis is over 13 million in 2008. From 2000-2005, the cost of treating allergic rhinitis is increasing from $6.1 billion to $11.2 billion with more than half of the money spent on prescription medications.
An allergen must be exposed to an organism to elicit a response, usually upon inhalation or ingestion. An allergen is a molecule, which causes an abnormal immunological response. Allergens tend to be protein but, can also be smaller and are called haptens.
Allergens can be bound by IgE immunoglobulins, B lymphocytes, T lymphocytes and Mast cells in the human. IgE-allergen immune complexes cause de-granulation of Mast cells and a release of histamines, locally and systemically. Histamine release can cause muscle cramps, muscle contraction, inflammation, erythema and edema of mucous membranes. Severe reactions can cause anaphylaxis and death.
Current diagnostic testing strategies are not cost effective, are time-intensive, are limited in the scope of antigen tested for, and/or are labor-intensive. Fast knowledge of individual health status allows for (i) appropriate medical treatment, (ii) the modification of behavior to decrease the likelihood disease incidence, morbidity, infection propagation and, (iii) the ability to save lives. The human genome project has identified 57 human genes coding for various cytochrome P450 enzymes that are involved in metabolic reactions. Cytochrome P450 isotypes and variants can interfere with drug bioactivity, drug metabolism and drug clearance and provide a confounding factor when prescribing drugs of vital importance to patients. The levels of cytochrome P450 isotypes are important factors to weigh when drugs with important side effects and drugs with narrow therapeutic windows are prescribed. There exists a need to quickly assess cytochrome P450 isotype levels in individual patients for appropriate drug selection with limited toxic effects. Thus there is also a need for genetic testing to detect cytochrome P450 isoform expression so that physicians can anticipate drug interactions in patients prior to prescribing medications.